This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. DEVELOPMENT OF ADVANCED MULTIDIMENSIONAL MICROSCOPY TECHNOLOGIES FOR MESOSCALE IMAGING The correlation of data acquired across scales and technologies is undertaken to reveal relationships among subcellular components, the detailed organization of constituent protein building blocks, and the functional role of each in normal and abnormal biological systems. The continued development and deployment of enhancements to the resource imaging instrumentation is critical to the improvement and further acceleration of our capability to investigate biomedical research questions, especially those that offer insight into structural and functional relationships across scales. To this end, we are maximizing the impact of our recently deployed 300KV energy filtering TEM/STEM instrument, integrating its advanced production capabilities and commissioning new imaging modes enabled by NCMIR-specified customizations. This team is also refining our existing IVEM and ultra-wide-field light microscopy resources to solidify the diverse techniques performed at this unique facility. We continue to develop and integrate new instrumentation technologies to bridge resolution gaps and augment our ability to conduct seamless multi-scale, multi-resolution microscopy--in particular serial blockface scanning electron microscopy. For each instrument, we focus on TR&D toincrease imaging throughput and fidelity and extend the fields-of-view achievable at high resolution. These activities balance engineering development with collaborative use and connect efforts to exploit new labeling and specimen-development technologies as well as image analysis and database utilization.